Creative Biostructure has focused on Nanodisc technology for years, our team has established an advanced Mempro™ detergent-solubilized system. Creative Biostructure is confident in providing high-quality membrane proteins reconstituted in Nanodisc services using detergent-solubilized system for either basic research or industrial applications.
The study of membrane mimetic which can keep the natural and functional activities remains challenging, on account of the requirement of extraction from their inartificial state and reconstituted in artificial milieus for the structure determination. while solubilizing in parallel the transmembrane segments with great spectral quality, only a elaborative use of a small number of mixed detergents did not disorder the cytoplasmic domain. Detergent-solubilized membrane proteins for structure measure by NMR spectroscopy or X-ray crystallography usually cut part of their function, for example, ligand binding and enzymatic activity accompanying structural changes.
Figure 1. The three stages of bilayer solubilization by detergents. (Biophysical journal 2013)
There is a method for reconstitution of membrane proteins in a bilayer-mimetic: self-assembling nanodiscs, beginning from a detergent-solubilized mixture containing all elements: target, lipid mixture of choice and membrane scaffold proteins(MSP). Besides, nanodiscs without detergents may fluctuate or unfold extra-membranous part of membrane proteins.
If the membrane protein under research is erratic to purification in detergent, or a set of the whole membrane proteins in a particular tissue are needed, combined into nanodiscs enables to be accomplished through direct solubilization of cell membranes by using detergents or membrane-active polymers. This approach makes the purification step to be neglected and improves the whole process of nanodisc self-assembly, which is occasionally critically essential for conservation of the native and functional form of the target membrane protein. To incorporate purified membrane proteins, the most crucial factor is efficient solubilization and holding back the aggregation of the target protein, these cases enable to be finished with excess lipid and scaffold protein along with keeping the correct overall stoichiometry. Factors that regulate the branch point leading to ideal incorporation of a target membrane protein contain the selection of detergent, the speed of detergent removal and the identities of the lipid and target.
As a pioneer and the undisputed global leader in Nanodisc technology, Creative Biostructure can perform Mempro™ detergent-solubilized system for membrane proteins reconstituted in Nanodisc services. Creative Biostructure provides other various Mempro™ Nanodisc reconstitution services. Please feel free to contact us for a detailed quote.
References: Lichtenberg D, Ahyayauch H, Goñi F M. The mechanism of detergent solubilization of lipid bilayers[J]. Biophysical journal, 2013, 105(2): 289-299.Tzitzilonis C, Eichmann C, Maslennikov I, et al. Detergent/nanodisc screening for high-resolution NMR studies of an integral membrane protein containing a cytoplasmic domain[J]. PLoS One, 2013, 8(1): e54378.Lichtenberg D, Ahyayauch H, Alonso A, et al. Detergent solubilization of lipid bilayers: a balance of driving forces[J]. Trends in biochemical sciences, 2013, 38(2): 85-93.Inagaki S, Ghirlando R, Grisshammer R. Biophysical characterization of membrane proteins in nanodiscs[J]. Methods, 2013, 59(3): 287-300.